Figure 5.
Figure 5. Flow cytometric analysis of EYFP expression in adult bone marrow cells. End-SCL-Cre-ERT;R26R-EYFP BM was harvested 6 weeks after 2 weeks of daily 2 mg tamoxifen (TAM) injections. (A) EYFP analysis within total adult BM. No Cre-mediated recombination was detected within BM of either untreated or in the majority of tamoxifen-treated mice. (B) EYFP expression within the c-kit+Sca-1+lineage- population. In 1 of 4 tamoxifen-treated mice, recombination was detected in total BM (A, right) as well as in the c-kit+Sca-1+lineage- population (B, right). (C) Additionally, we determined the activity of the end-SCL-Cre-ERT transgene in BM ECs (CD31+CD45-Ter119- population). In 2 of 3 mice, we could not detect any EYFP expression in BM endothelium. To demonstrate sufficient activity of the Rosa locus in the BM populations investigated, we analyzed CMV-Cre; R26R-EYFP BM as a positive control for EYFP expression. (B-C) Representative histograms (gray, end-SCL-Cre-ERT; R26R-EYFP; dotted, CMV-Cre; R26R-EYFP) are shown. Percentages indicate proportions of EYFP-positive cells of end-SCL-Cre-ERT; R26R-EYFP mice. FSC indicates forward light scatter.

Flow cytometric analysis of EYFP expression in adult bone marrow cells. End-SCL-Cre-ERT;R26R-EYFP BM was harvested 6 weeks after 2 weeks of daily 2 mg tamoxifen (TAM) injections. (A) EYFP analysis within total adult BM. No Cre-mediated recombination was detected within BM of either untreated or in the majority of tamoxifen-treated mice. (B) EYFP expression within the c-kit+Sca-1+lineage- population. In 1 of 4 tamoxifen-treated mice, recombination was detected in total BM (A, right) as well as in the c-kit+Sca-1+lineage- population (B, right). (C) Additionally, we determined the activity of the end-SCL-Cre-ERT transgene in BM ECs (CD31+CD45-Ter119- population). In 2 of 3 mice, we could not detect any EYFP expression in BM endothelium. To demonstrate sufficient activity of the Rosa locus in the BM populations investigated, we analyzed CMV-Cre; R26R-EYFP BM as a positive control for EYFP expression. (B-C) Representative histograms (gray, end-SCL-Cre-ERT; R26R-EYFP; dotted, CMV-Cre; R26R-EYFP) are shown. Percentages indicate proportions of EYFP-positive cells of end-SCL-Cre-ERT; R26R-EYFP mice. FSC indicates forward light scatter.

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